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猪SDHD基因的克隆、定位及其与胴体性状的关联研究

Cloning, mapping and association study with carcass traits of the porcine SDHD gene.

作者信息

Zhu Z M, Zhang J B, Li K, Zhao S H

机构信息

Department of Gene and Cell Engineering, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China.

出版信息

Anim Genet. 2005 Jun;36(3):191-5. doi: 10.1111/j.1365-2052.2005.01270.x.

Abstract

A 1320-bp cDNA containing the full coding region of the porcine succinate dehydrogenase complex, subunit D (SDHD) gene was obtained by random sequencing of clones from a Chinese Tongcheng pig 55-day fetal longissimus dorsi muscle cDNA library. Analysis of the SDHD gene across the INRA-University of Minnesota porcine radiation hybrid panel indicated close linkage with microsatellite marker SW2401, located on SSC9p21. The open reading frame of this cDNA covers 480 bp and encodes 159 amino acids. The deduced porcine amino acid sequence showed greater similarity with human and bovine protein sequences than with those from mouse and rat. The BLAST analysis of the porcine SDHD to NCBI identified Unigene Cluster Ssc.2586. Possible single nucleotide polymorphisms (SNP) were identified by alignment of expressed sequence tags in the cluster. The polymerase chain reaction (PCR) single strand conformation polymorphism, sequencing, and PCR restriction fragment length polymorphism were used to confirm and detect a synonymous polymorphic MboI site within the open-reading frame. Allele frequencies of this SNP were investigated in two commercial and five Chinese local pig breeds. These five Chinese breeds had very high frequencies for one allele, whereas frequencies of both alleles were intermediate in Large White and Duroc. An association analysis suggested that different SDHD genotypes have significant differences in loin-muscle area (P < 0.01).

摘要

通过对中国通城猪55日龄胎儿背最长肌cDNA文库中的克隆进行随机测序,获得了一个包含猪琥珀酸脱氢酶复合物D亚基(SDHD)基因完整编码区的1320 bp cDNA。对INRA - 明尼苏达大学猪辐射杂种细胞系面板中的SDHD基因进行分析,结果表明其与位于SSC9p21上的微卫星标记SW2401紧密连锁。该cDNA的开放阅读框覆盖480 bp,编码159个氨基酸。推导的猪氨基酸序列与人类和牛的蛋白质序列的相似性高于与小鼠和大鼠的序列。将猪SDHD与NCBI进行BLAST分析,确定了单基因簇Ssc.2586。通过比对该簇中的表达序列标签,鉴定出可能的单核苷酸多态性(SNP)。采用聚合酶链反应(PCR)单链构象多态性、测序和PCR限制性片段长度多态性来确认和检测开放阅读框内的一个同义多态性MboI位点。在两个商业猪品种和五个中国地方猪品种中研究了该SNP的等位基因频率。这五个中国品种中一个等位基因的频率非常高,而在大白猪和杜洛克猪中两个等位基因的频率均处于中等水平。关联分析表明,不同的SDHD基因型在腰大肌面积上存在显著差异(P < 0.01)。

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